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Biomedical subjects

S H Yap

Publications and source records attributed to S H Yap.

At least 109 records · Page 6Linked to original sources

Fine structure of the malaria parasite Plasmodium falciparum in human hepatocytes in vitro.

Recent advances in the ability to culture the hepatic forms of mammalian malaria parasites, particularly of the important human pathogen Plasmodium falciparum have provided novel opportunities to study the ultrastructural organisation of the parasite in its natural host cell the human hepatocyte. In this electron-microscopic and immunofluorescence study we have found the morphology of both parasite and host cell to be well preserved. The exoerythrocytic forms, which may be found at densities of up to 100/cm2, grow at rates comparable to that in vivo in the chimpanzee. In the multiplying 5- and 7-day schizogonic forms of the ultrastructural organisation of the parasite bears striking resemblances to other mammalian parasites, e.g., the secretory activity and distribution of the peripheral vacuole system, but also homology with avian parasites, e.g., in nuclear and nucleolar structure and mitochondrial form. The latter homologies support earlier suggestions of the close phylogenetic relationship of P. falciparum with the avian parasites. Evidence is also presented showing the persistence of the cytoskeleton of the invasive sporozoite within the cytoplasm of the ensuing rapidly growing vegetative parasites.

Animals↗

Changes in fluid-phase endocytosis in the rat with age and their relation to total albumin elimination.

Rates of fluid-phase endocytosis were determined in several organs and tissues of female WAG/Rij rats of several ages by using 125I-labelled polyvinylpyrrolidone ([125I]-PVP) as a marker. Liver, muscle and skin accounted for a high level of [125I]PVP uptake 28 h after injection. When PVP uptake was expressed per gram of organ/tissue, the liver and spleen were found to be the most active. An age-related increase in [125I]PVP uptake was seen at between 12 and 36 months of age in liver, kidneys and heart. Except for the kidneys this increase is caused by an increase in wet weight of these organs and not by an increase in the specific endocytic rate. These data, together with reported findings on the major sites of albumin catabolism, in liver, kidney, spleen and intestine, indicate that fluid-phase endocytosis is a main mechanism for the observed age-related increase in albumin elimination in these rats.

Aging↗

Activation of mutagens by hepatocytes and liver 9000 X g supernatant from human origin in the Salmonella typhimurium mutagenicity assay. Comparison with rat liver preparations.

The mutagenicity of 10 known genotoxic compounds, of several chemical classes, was measured in Salmonella typhimurium mutagenicity assays comprising isolated human hepatocytes or human liver 9000 X g supernatant (S9) from 4 different individuals, as activating system. The mutagenic activity of several compounds as determined with the Salmonella/hepatocyte suspension assay showed obvious differences when compared with the values obtained in the Salmonella/S9 plate assay. For instance, the mutagenic activity of BZ, DMN and DEN appeared to be much higher in the hepatocyte assay than in the S9 assay. However, 2-AF and 2-AAF were activated more effectively into mutagens in the S9 assay than in the hepatocyte assay. 2-AF was slightly more mutagenic than 2-AAF in the hepatocyte assay, whereas it was far more mutagenic than 2-AAF in the S9 assay. DMN was found more mutagenic than DEN in the hepatocyte assay, whereas in the S9 assay DEN appeared to be slightly more mutagenic. Furthermore, great interindividual differences in the metabolic activation of certain compounds, e.g. BZ and DMN, were observed in the hepatocyte suspension assay, whereas these variations were less evident in the S9 plate assay. Comparison of the mutagenicity data obtained with the human liver preparations, with those obtained with rat liver preparations, showed great interspecies differences in the capacity to activate certain chemicals into mutagens. The use of human liver preparations, in particular isolated human hepatocytes, may be of great value in studies on inter- and intraspecies variations in metabolic activation of genotoxic agents.

Animals↗

Mutagenicity and DNA-excision repair induced by isoniazid after metabolic activation by isolated human and rat hepatocytes.

The mutagenic potency of isoniazid (INH) (a widely used antitubercular drug) towards Salmonella typhimurium strain hisG46 was studied in the Salmonella/hepatocyte suspension-assay, comprising isolated human or rat hepatocytes as metabolic system. The potency of INH to induce DNA-excision repair in these hepatocytes was also measured. With rat hepatocytes, INH appeared to be only weakly mutagenic and did not induce significant increases in hepatocellular DNA-excision repair. With isolated hepatocytes of two human subjects, INH appeared also only weakly mutagenic. However, with hepatocytes of two other human subjects, INH was found to be highly mutagenic. Comparable results were obtained for the induction of hepatocellular DNA-excision repair.

Acetylation↗

Cryopreservation of adult human hepatocytes. The influence of deep freezing storage on the viability, cell seeding, survival, fine structures and albumin synthesis in primary cultures.

Isolated and cultured human hepatocytes provide a useful model for studies of the liver cell function in man. In vitro studies using human hepatocytes are scarce, due to the limited availability and the lack of suitable methods for storage. In this study, we report the effect of deep freezing storage on the viability, fine structures and albumin synthesis of human adult hepatocytes in classical culture conditions. Hepatocytes were isolated using collagenase perfusion (9 isolations). The cell yield was 4-37 X 10(8) with a viability of 60-87%. Cryopreservation was performed in medium containing 10% DMSO and 20% fetal calf serum using a Cryoson BV-4 programmable freezer (0 degree C for 5 min, followed by a freezing rate of 1.5 degrees C/min for 20 min and 7 degrees C/min for 10 min). The cells were stored for 25-275 days in the liquid nitrogen vapor phase (-150 degrees C). Within 16 h about 80% of viable cells from freshly isolated hepatocytes whereas after cryopreservation, 55% of viable cells as determined by Trypan Blue exclusion before the cryopreservation attached to plastic and survived. Electron microscopy showed well developed tight junctions, structures similar to bile canaliculi. Cell polarity was evident. However, 'bleb' formation, more lipid droplets and lysosomes were found in cryopreserved hepatocytes during a short period after thawing. At the 3rd week, cells detached and died. These changes were associated with increased secretion of lactate dehydrogenase, whereas the albumin secretion dropped (from 10 to 4 micrograms/micrograms DNA), regardless of whether hepatocytes were cultured from fresh preparations or after cryopreservation. These findings suggest the cryopreservation is a useful technique to preserve hepatocytes for in vitro studies. Nevertheless, an improved method is necessary to increase the efficiency of cell seeding after cryopreservation.

Albumins↗

[Gastric polyps and their relations with gastric carcinoma].

In 31 of 33 patients with polyps of the stomach who underwent gastroscopy and x-ray examination the polyps could be diagnosed roentgenologically. 11 patients had multiple polyps, which were in 8 cases hyperplastic polyps, in 3 "other polyps", whereas in 20 patients with solitary polyps 11 cases showed hyperplastic, in 5 cases adenomatous and in 4 cases "other" polyps. After x-ray analysis it can be stated that in our cases the premalignant adenomatous polyp always occurred solitary.

Adult↗

Pancreatic mass due to Strongyloides stercoralis infection: an unusual manifestation.

A 30-year-old man returning from a trip through India complained of epigastric pain, diarrhea, loss of weight, and jaundice. Stool examination was positive for Strongyloides stercoralis. On ultrasonographic investigation, intravenous cholangiography, and endoscopic retrograde cholangiopancreaticography, stenosis of the distal common bile duct and enlargement of the pancreatic head were observed. The patient was treated for the Strongyloides infection. The jaundice disappeared within days, the laboratory finding normalized, and the patient recovered rapidly. However, the ultrasonographic examination showed a complete recovery in the course of about 2 years. From this report we conclude that in patients with obstructive jaundice and Strongyloides stercoralis infection, serious abnormalities resembling pancreatic tumour or pancreatitis can be found on ultrasonographic and radiological investigations, and this should be considered in the differential diagnosis of pancreatic mass in patients living in or returning from endemic areas.

Adult↗

[Choledocholithiasis in a gallbladder without stones].

In 4 out of 190 patients, who underwent a cholecystectomy, stones were only found in the common bile duct and none in the gallbladder. The radiological data of these 4 patients were retrospectively analysed. In 2/3 patients sonography showed a dilatation of the common bile duct, whereas stones in the common bile duct were shown by IVC in 2/2 patients, ERCP in 3/3 patients and PTC in 1/1 patients. In 1/3 patients also, in whom an ERCP had been performed, stones were suspected in the gallbladder, but could not be verified during cholecystectomy.

Adult↗

Fibrinogen fragments X, Y, D and E increase levels of plasma fibrinogen and liver mRNAs coding for fibrinogen polypeptides in rats.

Previously, we demonstrated that in vivo regulation of liver fibrinogen synthesis occurs via the fibrinogen mRNA level. However, the molecular regulatory mechanism of fibrinogen synthesis is still not well understood. Fibrinogen or fibrin degradation products might play an important role in regulating fibrinogen synthesis. In our present study, we have injected rats intraperitoneally with purified homologous fragments and measured the liver content of mRNA specific coding for fibrinogen. Increased levels of fibrinogen mRNA and elevated plasma fibrinogen concentrations were observed in rats after administration of fibrinogen degradation products X, Y, DEGTA, Dcate or E. Fragment E or E' has a less stimulatory effect than X, Y or Dcate, whereas cross-linked fibrin degradation product D dimer does not increase fibrinogen synthesis. This article reports for the first time a stimulatory effect of the high molecular weight fibrinogen degradation products on fibrinogen synthesis.

Animals↗

The influence of glucocorticoid on albumin synthesis and its messenger RNA in rat in vivo and in hepatocyte suspension culture.

Corticosteroids are known to stimulate the synthesis of a number of liver-specific proteins. The reports regarding the effect of glucocorticoid on albumin synthesis in vivo and in vitro are controversial. In an attempt to determine the mechanism by which glucocorticoid exerts its influence on hepatic albumin synthesis and to find an explanation for the conflicting data, we have studied the effect of dexamethasone disodium phosphate on albumin synthesis and albumin messenger RNA as determined by the molecular hybridization technique in hepatocytes in rat in vivo and in suspension culture. In hepatocyte suspension culture, addition of 0.48 microM dexamethasone in medium at zero time led to a significant increase (20%) in incorporation of labeled precursor into albumin as compared to control experiments; this was accompanied by a maintainance of the initial level of full-length albumin mRNA for a 9 h period. In hepatocytes cultured without dexamethasone in the medium there was a progressive loss of albumin mRNA content. Despite this finding, dexamethasone was not able to increase the albumin mRNA content in hepatocyte to a level higher than the initial value. Moreover, administration of this hormone either intraperitoneally or intravenously into rats did not lead to enhanced cell-free albumin synthesis or to an increased level of albumin mRNA. These findings suggest that glucocorticoid does not play an essential role in the regulation of albumin synthesis in vivo. In vitro, however, glucocorticoid leads to a preservation of the initial level of albumin mRNA and thus plays a role in the control of spontaneous dedifferentiation of liver cells in culture.

Albumins↗

Detection of hepatitis B virus DNA in serum and relation with the IgM class anti-HBc titers in hepatitis B virus infection.

Sera from four groups of patients wtih different serologic markers of HBV infection were examined for HBV DNA using molecular hybridization technique and for IgM class anti-HBc using an ELISA based on the antibody capture principle. Results of HBV DNA assay were generally in good agreement with the presence of HBeAg. However, HBV DNA was found in 13% of anti-HBe+ sera and in one patient with anti-HBc as a sole marker. IgM anti-HBc was detected at high titers in acute hepatitis B patients and was also present during the "window-period." This marker was also found, though less frequently when other markers for HBV infectivity were absent, in chronic hepatitis B patients and healthy carriers. From these findings we conclude that the HBV DNA assay provides a reliable method of detecting the infectious agent, particularly in anti-HBe+ sera and sera with anti-HBc as a sole marker. The assay for IgM anti-HBc is useful for establishing the diagnosis of recent infection in patient with anti-HBc as a sole marker, and during acute hepatitis with very high aminotransferase values, a condition in which HBV DNA may be undetectable.

Acute Disease↗

Absence of detectable hepatitis B virus DNA in sera and liver of chimpanzees with non-A, non-B hepatitis.

The risk of hepatitis B infections has been reduced by screening of blood donors for hepatitis B surface antigen (HBsAg). However, recipients remain at significant risk of developing post-transfusion hepatitis. Studies have shown that non-A, non-B hepatitis virus(es) are responsible for the majority of post-transfusion hepatitis infections. In spite of many efforts, these non-A, non-B hepatitis viruses have not yet been identified. Epidemiological studies, however, suggest that non-A, non-B hepatitis shares many features with hepatitis B. Recently, Wands et al [1982] showed, in chimpanzees infected with non-A, non-B hepatitis agents, the presence of antigenemia or viremia by radioimmunoassay with monoclonal antibodies directed toward distinct determinants of HBsAg and by molecular hybridization analysis. They suggested that non-A, non-B hepatitis agents may be related, but distinct variant(s) of hepatitis B virus (HBV). In this study, five chimpanzees were inoculated with three different agents that have been shown to transmit non-A, non-B hepatitis. The following inocula were used (I) a factor VIII preparation kindly provided by D.W. Bradley, (II) acute phase serum from a chimpanzee infected with the F strain kindly provided by A.J. Zuckerman, and (III) a DS-antigen serum previously shown by us to transmit non-A, non-B hepatitis [Duermeyer et al, 1983]. All chimpanzees developed a rise in transaminase levels between 8 and 10 weeks after inoculation. None of the chimpanzees was positive for any markers of HBV infection. No evidence was obtained of infection with hepatitis A, cytomegalovirus, or Epstein-Barr virus. One chimpanzee developed chronic liver disease.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Phase II trial of 5-fluorouracil, adriamycin and cisplatin (FAP) in advanced gastric cancer.

Twenty patients (15 male, 5 female) with nonresectable gastric adenocarcinoma were treated with FAP (5-fluorouracil 300 mg/m2 IV on days 1-5, adriamycin 50 mg/m2 IV on day 1, cisplatin 20 mg/m2 IV on days 1-5). Each course was repeated every 21 days. Eighteen patients were evaluable for response. The median age was 51 years, the range extending from 34 to 68. None had undergone chemotherapy. The median Karnofsky performance score was 80%. Nine (50%) partial responses (PR) and eight (44%) cases of stable disease (SD) were observed. One patient showed progression of the disease and died after 6 months. The median duration of response was 6+ months for PR and 6 months for SD. The median survival was 12 months. FAP toxicity was moderate, with the median WBC nadir 3.2 X 10(9)/l (range 0.7-4.2). One patient in PR died of septicemia. Nausea and vomiting were not dose-limiting. Neuropathy was mild in four and moderate in two patients. This FAP combination appears to be as effective with respect to response rate and duration as reported for 5-fluorouracil, adriamycin and mitomycin C (FAM).

Adenocarcinoma↗

Roentgenologic changes of the colon in Campylobacter infection.

A double-contrast examination of the colon was performed during diarrheal illness in 5 patients with Campylobacter infection. The findings in these 5 patients were aphthoid ulcers and stippled appearance. In 1 patient with stool culture positive for Campylobacter, double-contrast study of the colon 9 days later showed no abnormalities. In a 12-year-old boy lymphoid hyperplasia could be observed as well as the aphthoid ulcers. All the abnormalities were segmental, more often localized in the rectosigmoid. No changes were found in the ascending colon. Histopathologic studies of the biopsy specimens taken endoscopically in 3 patients showed a nonspecific inflammatory reaction.

Adult↗

Mutagenicity of five arylamines after metabolic activation with isolated dog and human hepatocytes.

The mutagenicity of benzidine (BZ) N-acetylbenzidine (MABZ), N,N'-diacetylbenzidine (DABZ), 4-aminobiphenyl (4-AB) and 2-aminoanthracene (2-AA) towards Salmonella typhimurium strain TA1538 was measured in the presence of isolated hepatocytes from dog and man. The influence of paraoxon, an inhibitor of the deacetylation reaction, on the mutagenicity of these compounds was also investigated. Obvious interspecies differences in the mutagenic activation of benzidine and its acetylated-derivatives were seen. However, with liver cell preparations from both species it was found that MABZ and DABZ were more mutagenic than BZ itself. 4-AB appeared to be weakly mutagenic in the presence of human hepatocytes but non-mutagenic with dog hepatocytes. 2-AA was highly mutagenic in both species. When human hepatocytes were used as the metabolic factor, the mutagenicity of all arylamines decreased in the presence of paraoxon. With dog hepatocytes, however, the mutagenicity of all arylamines except DABZ was enhanced in the presence of paraoxon.

Adult↗